Facile preparation of carbon nanotube-conducting polymer network for sensitive electrochemical immunoassay of Hepatitis B surface antigen in serum

A novel electrochemical immunosensor built on three dimensional carbon nanotube-conducting polymer (CNT-CP) network is reported for detection of Hepatitis B surface antigen (HBsAg) in human serum. The CNT-CP network is prepared by drop-drying of CNT solution on glassy carbon electrode, followed by electrochemical polymerization of poly (pyrrole propionic acid) (pPPA) film to crosslink and stabilize the CNTs, wherein the CNTs form the backbone of the network, and offer great specific surface areas for antibody attachment, and confer good conductivity for electrochemical detection, while the conducting film integrates the carbon nanotubes into a stable network due to its self-limiting growth behavior and provides abundant carboxyl groups for covalent immobilization of probe proteins. As a unique matrix, the CNT-CP network enables sensitive electrochemical detection of HBsAg biomarker by using alkaline phosphatase (ALP)-conjugated secondary antibodies under sandwich format coupling with the ALP substrate solution, p-aminophenyl phosphate (PAPP), reaching a detection limit of 0.01 ng/mL with a dynamic range of 5 orders of magnitude. (C) 2011 Elsevier B.V. All rights reserved.